I will be using amicon filter for the first time to conc. my lentiviral particles produced. The total weight of the plasmid DNA used to transfect accounts to 40.7kb. How do i calculate my produced viral particle size from it and then select MWCO accordingly. plz help. Also what type of membrane should i select as this lentiviral soup contains pluripotency proteins. plz help.
Hi there,
It's a matter of pore diameter, not MWCO. Lentivirus particles generally exhibit a diameter between 80 and 100nm. The suitable membrane for concentrating such material is membrane with 300K MWCO (estimated pore size is 30nm). See below for more info:
https://cellculturedish.com/questions/what-mwco-and-membrane-is-best-suited-for-viral-vector-concentration-such-as-lentivirus-or-aav/
In the Addgene protocol for pLKO.1(lentivirus plasmid) it suggests to use a 0.45um filter to remove the cells. If you use a 0.2um filter it says it is likely to shear the envelope of your virus. So I would be sure that the filter you are using to concentrate your lentivirus particles will not shear your virus.
Agree with Dominique Liger here - MWCO values on centricon packings are actually estimated for globular particles, so one should be careful when working with molecules of different shape.
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